ORIGINAL PAPER

Differential acclimation capacity to frost in sugarcanevarieties grown under field conditions

Philippus D. R. van Heerden

Received: 11 June 2013 / Accepted: 24 September 2013 / Published online: 29 September 2013

� Springer Science+Business Media Dordrecht 2013

Abstract In certain sugarcane (Saccharum spp. hybrids)

production regions of the world, including South Africa,

frost frequently occurs. Great demand exists for frost tol-

erant sugarcane varieties as production in these areas could

become more profitable. Two Louisiana (USA) varieties,

known to yield high sugar even when immature (LCP

85-384 and HOCP 96-540), and two South African varie-

ties (N21 and N36) were evaluated in a field trial for frost

tolerance. O–J–I–P chlorophyll a fluorescence transients

were recorded in youngest fully-expanded leaves of these

varieties on several occasions before and following expo-

sure to frost. Analysis of these transients revealed that

varieties N36 and LCP 85-384 were capable of cold

acclimation following the first frost, while N21 and HOCP

96-540 lacked similar capability. Exposure to further frosts

altered the fluorescence transients in a variety-specific

fashion, with recovery in N36 and LCP 85-384 towards

baseline kinetics, but with further deterioration in N21 and

HOCP 96-540. Between the first frost and harvest, esti-

mated recoverable crystal (ERC) content values in cane

stalks of N36 and LCP 85-384 increased by 26 and 20 %

respectively, while in N21 and HOCP 96-540 ERC content

values only increased by 8 and 11 % respectively. Conse-

quently, N36 and LCP 85-384 ultimately achieved the

highest ERC yields (tons/hectare). The ability to maintain

high ERC accumulation capacity for longer following frost

could be an important factor determining sugarcane yield

performance in frost-prone areas. In addition, O–J–I–P

fluorescence rise kinetics show promise as a rapid screen-

ing tool for assessment of cold acclimation potential in

sugarcane.

Keywords Chlorophyll a fluorescence � Cold

acclimation � Frost tolerance � Sucrose accumulation �Sugarcane

Abbreviations

ERC Estimated recoverable crystal

NADP-MDH NADP-malate dehydrogenase

PIABS Performance index

Introduction

Due to their mostly tropical and sub-tropical distribution,

C4 plants generally are more cold-sensitive than C3 plants

(Berry and Bjorkman 1980). Cold tolerance also differs

between the three C4 subtypes, with those species

belonging to the NADP-malic enzyme (NADP-ME) sub-

type generally being the most cold-sensitive (Edwards et al.

1985). In a recent study the effects of low temperatures on

C4 and C3 sub-tropical grasses were compared in Alloter-

opsis semialata, a unique species with both C4 and C3

subspecies co-occurring in sub-tropical parts of South

Africa (Osborne et al. 2008). These authors found high

levels of freezing stress injury in leaves of both sub-species

in the unhardened state. However, a prolonged chilling

treatment introduced as a hardening treatment before the

freezing stress, reduced injury in the C3 subspecies by

50 %, but not in the C4 subspecies. It was concluded that

the C4 subtype lacked the ability for cold acclimation

P. D. R. van Heerden (&)

South African Sugarcane Research Institute,

Mount Edgecombe 4300, South Africa

e-mail: riekert.vanheerden@sugar.org.za

P. D. R. van Heerden

Department of Plant Production and Soil Science,

University of Pretoria, Pretoria 0028, South Africa

123

Plant Growth Regul (2014) 72:181–187

DOI 10.1007/s10725-013-9850-3

(Osborne et al. 2008). However, leaves of some C4 grass

species can resist freezing injury at temperatures of less

than -10 �C (Sage and Kubien 2007). Therefore, although

tolerance to freezing is not completely absent, most C4

grasses do not demonstrate significant cold acclimation

capability, and are regarded as sensitive to freezing stress

(Rowley et al. 1975; Rowley 1976).

As a consequence of its recognised sensitivity to low

temperature, cultivation of sugarcane in frost-prone areas

across the world often necessitates shorter growing cycles

in order to escape the detrimental after-effects of frost

damage such as leaf and apical meristem mortality, profuse

side-shooting and cane deterioration (Eggleston et al.

2004). Frost is very common in the Midlands region of

Kwazulu-Natal (South Africa), especially in valley bot-

toms. Harvesting sugarcane annually in the Midlands

region is less profitable because the full yield potential of

the crop, which is usually attained after 18–24 months, is

not yet realised after only 12 months of growth. Hence,

there is urgent need for identifying frost tolerant varieties,

or varieties yielding high sugar even when harvested

young, to increase profitability in frost affected areas.

Low temperatures have been shown to negatively affect

photosynthesis in sugarcane, in particular in the form of less

efficient energy transfer to reaction centres and slower

reduction of the primary electron acceptors of photosystem

II (PSII) (Ebrahim et al. 1998). In addition, marked reduc-

tion in the activities of NADP-malate dehydrogenase

(NADP-MDH) and pyruvate, orthophosphate dikinase

(PPDK) have also been reported in sugarcane exposed to

low temperature (Du et al. 1999). These authors proposed

that the inhibition of NADP-MDH and PPDK activity

limited CO2 assimilation capacity with subsequent pho-

toinhibition and upstream damage to the electron transport

systems. These results suggest that electron transport

capacity is a main target during low temperature stress and

that chlorophyll a fluorescence might be a sensitive tool for

probing the degree of damage and acclimation responses in

sugarcane. Although we are not aware of any studies in

sugarcane, chlorophyll a fluorescence has been used suc-

cessfully in the past to assess the effects of freezing stress

and subsequent recovery in winter wheat cultivars (Rapacz

2007; Rapacz et al. 2011). However, a limitation of these,

and numerous other studies using chlorophyll a fluores-

cence to probe stress effects in crop plants, is that the

observed effects is very seldom related to actual impact on

yield. Many of these studies are typically also conducted

under controlled growth conditions in potted plants, where

it is extremely difficult to realistically mimic impacts of

stress factors on yield. Moreover, it has also been suggested

that controlled growth environments might often introduce

artefacts, which then produces an unrealistic picture of cold

acclimation processes (Trischuk et al. 2006).

The research reported in this paper addressed these

limitations and lack of current understanding by perform-

ing field evaluation of the response of different sugarcane

varieties to frost. A further novel aspect of the work was

the subsequent relating of these responses to sugar accu-

mulation and yield. This study was premised on the

hypothesis that varying degrees of cold acclimation

capacity exists in sugarcane and that field evaluation

involving chlorophyll a fluorescence measurements would

be a suitable approach to investigate these responses and

subsequent impacts on yield.

Materials and methods

Variety selection

Two quick-maturing imported sugarcane varieties (LCP

85-384 and HOCP 96-540) from Louisiana (USA), known

to yield high sugar even at a relatively young age of only

8–9 months (Dufrene and Tew 2004; Eggleston et al.

2004), and two well-known South African varieties (N21

and N36) were included in this study. All varieties were

propagated using the NovaCane� tissue culture protocol

(Snyman et al. 2008) to ensure disease-free material for

field planting.

Field trial details

A field trial was established in a frost-prone valley-bottom

on a commercial farm near the town of New Hanover in the

Midlands-North region of Kwazulu-Natal, South Africa.

Trial plots of sufficient area to accommodate five sugarcane

rows 10 m long spaced 1 m apart were marked out. Soil

samples from 0 to 20 cm depth were collected from each of

the plots and subjected to chemical and soil texture analy-

ses. The soil characteristics of each plot were then used with

Principal Component Analysis (ADE-4 software) to map

the variability across the trial area. Relationships between

plots were examined with cluster analysis and individual

plots assigned to five replicates per variety in a way that

would minimise potential effects of soil variability. The

trial was planted on 18 October 2007. Trial management by

the farmer was done as per usual commercial practice. The

first light frost occurred during mid-June 2008 and was

followed by a series of heavier frosts between 26 June and 3

July 2008. The heaviest frost occurred on 11 July 2008. The

trial was harvested on 23 July 2008.

Chlorophyll a fluorescence measurements

For a physiological assessment of frost effects on the

varieties O–J–I–P chlorophyll a fluorescence transients

182 Plant Growth Regul (2014) 72:181–187

123

(Strasser and Govindjee 1992) were recorded in dark

adapted youngest fully-expanded leaves. These measure-

ments were started on 25 April 2008, approximately

6 weeks before the first frost and were continued until

shortly before harvest. Measurements were repeated

approximately every 2 weeks, which included a measure-

ment a few days before the first frost, a few days after the

first frost and also after several additional frosts. On each

occasion measurements were done on the broadest mid-

section of the youngest fully expanded leaves of four

randomly-selected plants per replicate plot. In total 20

plants per variety were measured on each occasion. Mea-

surements were conducted with a PEA fluorescence meter

(Hansatech Instruments Ltd., King’s Lynn, Norfolk, PE 30

4NE, UK). The transients were induced by a red light (peak

at 650 nm) of 3,200 lmol m-2 s-1 provided by the PEA

instrument through an array of six light-emitting diodes.

The JIP-test (Strasser and Strasser 1995) was subse-

quently employed to analyse each recorded transient. The

following data from the original measurements were used:

maximal fluorescence intensity (FM); fluorescence intensity

at 50 ls (considered as F0); fluorescence intensity at

300 ls (F300 ls) required for calculation of the initial slope

(M0) of the relative variable fluorescence (V) kinetics; the

fluorescence intensity at 2 ms (the J step) denoted as FJ. VJ

was calculated as (FJ - F0)/(FM - F0). The data was used

to calculate (Eq. 1) the Performance index (PIABS), which

is a sensitive indicator of electron transport efficiency

(Kruger et al. 1997; Strasser et al. 2000; Force et al. 2003;

Strauss et al. 2007).

PIABS ¼1� ðF0=FMÞ

M0=VJ

� FM � F0

F0

� 1� VJ

VJ

ð1Þ

Yield parameters

A 12-stalk sample was taken from each plot just prior to the

first frost to determine a range of milling quality charac-

teristics, including estimated recoverable crystal (ERC)

content, which is an estimate of the recoverable value of

sugarcane delivered to the sugar mill. The ERC content

(%) was calculated per stalk fresh weight (FW) as (Eq. 2):

ERC content %ð Þ ¼ aS� bN� cF ð2Þ

where S = sucrose content (%) per stalk FW; N = non-

sucrose content (%) per stalk FW; F = fibre content per

stalk FW and a, b, c are industry determined factors

representing the loss of each component (0.978, 0.535, and

0.018 respectively) during sucrose extraction at the mill. At

harvest, the three centre cane rows in each plot were cut

and bundled by hand and weighed using a hydraulic grab

apparatus equipped with a load cell to determine cane yield

(tons) per hectare (ha). Another 12-stalk sample was taken

from each plot for calculation of ERC content at harvest

(Eq. 2). The ERC yield (t/ha) per plot was subsequently

estimated as (Eq. 3):

ERC yield t=hað Þ ¼ ERC content� Cane yield ð3Þ

Statistical analysis

All the variables were analysed using one-way ANOVA.

Each variable was first tested for normality and homoge-

neity using the Shapiro–Wilk and Bartlett tests respectively

(Genstat v.14). The post hoc test used was the least sig-

nificant difference (LSD) (5 %) test.

Results

Effects of frost on the performance index (PIABS)

In Fig. 1a, b the Performance index (PIABS) values are

shown for the four varieties recorded from 6 weeks before

the first frost (first four data points including one marked

with black arrow), 5 days after the first frost (data point

marked with dark-grey arrow), after several subsequent

frosts (data point marked with light-grey arrow) and until

shortly before harvest. All varieties showed a decline in

PIABS values following the first light frost event (decline in

PIABS values from black to dark-grey arrow). This decline

in PIABS values (relative to pre-frost values marked by

black arrows) were 20, 30, 14 and 26 % in N36, LCP

85-384, N21 and HOCP 96-540 respectively. However,

following additional frost events (time period indicated

between dark-grey and light-grey arrows) N36 and LCP

85-384 maintained PIABS values at similar levels for at

least 2 weeks longer (Fig. 1a) than the other two varieties.

In N21 and HOCP 96-540 these additional frosts led to a

large decline in PIABS values (Fig. 1b). At this point in time

(marked by light-grey arrows) the decline in PIABS values

(relative to pre-frost values marked by black arrows) was

16, 32, 48 and 57 % in N36, LCP 85-384, N21 and HOCP

96-540 respectively clearly illustrating the additional

decline that occurred in N21 and HOCP 96-540. Ulti-

mately, the most severe frost event that occurred just prior

to harvest led to severe damage in all varieties.

Effects of frost on O–J–I–P fluorescence rise kinetics

Illumination of the dark-adapted leaves induced charac-

teristic changes in the intensity of chlorophyll a fluores-

cence, known as the Kautsky effect (Kautsky and Hirsch

1931). When the Kautsky transient is plotted on a loga-

rithmic time scale the fluorescence rise kinetics are poly-

phasic, clearly exhibiting two intermediate steps known as

J and I (Strasser and Govindjee 1992), hence the notation

O–J–I–P for the fast rise of the fluorescence transient. In

Plant Growth Regul (2014) 72:181–187 183

123

Fig. 2 O–J–I–P fluorescence transients recorded in fully

expanded leaves of the four varieties a few days before the

first frost (5 June 2008), a few days after the first frost (20

June 2008), and also after several additional frosts (3 July

2008) are shown. The transients were normalized at the

O (F0) and P (FM) steps in order to reveal changes in the

shape of the transients between these two extremes more

clearly. In all four varieties the frost-induced changes in the

shape of these transients are clearly visible.

The variety-specific effects of frost on the O–J–I–P

fluorescence rise kinetics were visualised by the construc-

tion of difference in variable fluorescence (DV) curves as

detailed by Strauss et al. (2007). For the construction of DV

curves the normalised values (Fig. 2) derived from tran-

sients recorded after the first light frost (20 June 2008), and

after several subsequent frosts (3 July 2008), were sub-

tracted from the normalised values derived from transients

recorded prior to the first frost (5 June 2008). The black

lines in Fig. 3 represents the result (zero values) obtained

after subtraction of the normalised values derived from

transients recorded prior to the first frost (5 June 2008)

from itself in order to produce a zero baseline. Any devi-

ation (from the black zero baseline) in DV curves derived

after the first light frost (light-blue curves), and after sev-

eral subsequent frosts (dark-blue curves) thus illustrates the

degree of alteration in O–J–I–P fluorescence rise kinetics

induced by frost in the four varieties (Fig. 3).

Construction of these DV curves, using the pre-frost

O–J–I–P fluorescence rise kinetics (5 June 2008, black

horizontal lines) for each variety as a baseline, revealed that

the first frost (20 June 2008, light-blue curves) induced a

perturbation in kinetics of very similar shape and magnitude

in all four varieties (Fig. 3). However, after several sub-

sequent frosts (3 July 2008, dark-blue curves) the kinetics

were altered in a variety-specific fashion, with clear

recovery in N36 and LCP 85-384 towards baseline kinetics

(downward pointing green arrows), but with further dete-

rioration in N21 and HOCP 96-540 (upward pointing green

arrows), with the peaking of deviation located at 2 ms (J-

step). These changes in O–J–I–P fluorescence rise kinetics

corresponded well with the changes in PIABS values where

all four varieties experienced a decline following the first

light frost (Fig. 1, dark-grey arrows and Fig. 3, light-blue

curves) but where N36 and LCP 85-384 then stabilised

PIABS values at similar levels for at least 2 weeks longer

while N21 and HOCP 96-540 experienced further declines

(Fig. 1, light-grey arrows and Fig. 3, dark-blue curves).

Effects of frost on yield parameters

Samples were taken from each treatment plot just prior to

the first frost, and again at harvest, to determine the effect

of frost on accumulation of ERC in the stalks of the four

varieties. These results showed that N36 and LCP 85-384

increased ERC content (%) by 26 and 20 % between the

pre-frost state and harvest, compared to N21 and HOCP

96-540, where the increases were only 8 and 11 % (Fig. 4).

In the case of N36 and LCP 85-384 the increases in ERC

content were statistically significant, while the much

smaller increases in N21 and HOCP 96-540 were not.

Although N36 and N21 had similar cane yields at har-

vest (results not shown), N36 had 30 % higher ERC yields

(t/ha) than N21 (Fig. 5). Likewise, the reported ERC yield

advantage of HOCP 96-540 over LCP 85-384, that are

usually observed under normal growing conditions (Duf-

rene and Tew 2004), was reversed following frost.

Discussion

The O–J–I–P fluorescence transient reflects the filling up of

the electron acceptor side of PSII (QA, QB and PQ pool)

Per

form

ance

inde

x (P

I AB

S)

0

5

10

15

20

N36 LCP 85-384

Date of measurement

25/04

/2008

08/05

/2008

23/05

/2008

05/06

/2008

20/06

/2008

03/07

/2008

18/07

/2008

Per

form

ance

inde

x (P

I AB

S)

0

5

10

15

20

N21 HOCP 96-540

A

B

Fig. 1 The effect of frost on Performance Index (PIABS) values in the

sugarcane varieties N36, LCP 85-384 (a) and N21, HOCP 96-540 (b).

Measurements were taken from 6 weeks before the first frost (first

four data points including one marked with black arrow), 5 days after

the first frost (dark-grey arrow), after several subsequent frosts (light-

grey arrow) and until shortly before harvest

184 Plant Growth Regul (2014) 72:181–187

123

with electrons from the donor side of PSII (Papageorgio

1975; Lavorel and Etienne 1977; Strasser and Govindjee

1992). The relationship of these events to the O–J–I–P

fluorescence transient was suggested by Strasser et al.

(1995) to be the following: O, minimal Chl a fluorescence

yield (highest yield of photochemistry); O to J, reduction of

QA to QA- (photochemical phase, light intensity dependent);

J to I to P, reduction of the PQ pool (non-photochemical

phase). Since the O–J–I–P fluorescence transient reflects

the kinetics and heterogeneity involved in the filling up of

the PQ pool with electrons, it can be used as a sensitive tool

to investigate the photosynthetic apparatus in vivo (Strasser

et al. 1995). The shape of the O–J–I–P fluorescence tran-

sient has been found to be very sensitive to various types of

stress (Van Rensburg et al. 1996; Kruger et al. 1997; Lazar

and Ilık 1997; Tsimilli-Michael et al. 1999). From the data

shown in Figs. 2 and 3 two important observations

regarding the effect of frost on the O–J–I–P fluorescence

transients in the four sugarcane varieties can be made:

(a) the shape of the O–J–I–P fluorescence transients

recorded in leaves of the varieties following frost exposure

differed from those recorded in the leaves prior to the first

N36

Flu

ore

scen

ce (

mV

)

0.0

0.2

0.4

0.6

0.8

1.0

5 June 2008 20 June 2008 3 July 2008

LCP 85-384

Flu

orescen

ce(mV

)

0.0

0.2

0.4

0.6

0.8

1.0

5 June 2008 20 June 2008 3 July 2008

N21

Time (ms)

0.1 1 10

Flu

ore

scen

ce (

mV

)

0.0

0.2

0.4

0.6

0.8

1.0

5 June 2008 20 June 2008 3 July 2008

HOCP 96-540

Time (ms)

0.1 1 10100 1000 100 1000

Flu

orescen

ce (mV

)

0.0

0.2

0.4

0.6

0.8

1.0

5 June 2008 20 June 2008 3 July 2008

O

J

IP

Fig. 2 Effects of dark chilling

on O–J–I–P fluorescence

transients (normalised between

O and P) recorded in fully

expanded leaves of the four

sugarcane varieties. Transients

were recorded pre-frost (5 June,

black curves), after the first frost

(20 June, light-blue curves) and

again after several additional

mild frosts (3 July, dark-blue

curves). The position of the

main steps (O, J, I and P) is

indicated in the top left-hand

figure

N36

Flu

ore

scen

ce (

mV

)

-0.04

0.00

0.04

0.08

0.12

0.165 June 200820 June 20083 July 2008

LCP 85-384

Flu

orescen

ce (mV

)

-0.04

0.00

0.04

0.08

0.12

0.165 June 200820 June 20083 July 2008

N21

Time (ms)

0.1 1 10 100 1000

Flu

ore

scen

ce (

mV

)

-0.04

0.00

0.04

0.08

0.12

0.165 June 200820 June 20083 July 2008

HOCP 96-540

Time (ms)

0.1 1 10 100 1000

Flu

orescen

ce (mV

)

-0.04

0.00

0.04

0.08

0.12

0.165 June 200820 June 20083 July 2008

Fig. 3 Difference in variable

fluorescence (DV) curves

constructed by subtraction of

normalised (O–P) fluorescence

values recorded pre-frost

(5 June, black horizontal

baseline) from those recorded

after the first frost (20 June,

light-blue curves) or after

several additional mild frosts

(3 July, dark-blue curves)

Plant Growth Regul (2014) 72:181–187 185

123

frost, and (b) a conspicuous difference existed between the

four varieties with regard to the extent by which the shape

of the transients was altered by frost, especially at the

J-peak (2 ms). An increase in fluorescence intensity at

2 ms is usually interpreted as evidence of accumulation of

the fraction of reduced QA- pool (Strasser et al. 1995; Lazar

and Ilık 1997), possibly due to a decrease in electron

transport beyond QA- (Haldimann and Strasser 1999).

Construction of DV curves, using the pre-frost O–J–I–P

fluorescence rise kinetics for each variety as a baseline,

revealed that the first frost induced a perturbation in

kinetics of very similar shape and magnitude in all four

varieties. However, after further frosts the kinetics were

altered in a variety-specific fashion, with clear recovery in

N36 and LCP 85-384 towards baseline kinetics. These

results depict a typical example of stress acclimation where

exposure to a light frost hardens the plants to better tolerate

subsequent heavier frost. These results suggest that N36

and LCP 85-384 were capable of cold acclimation fol-

lowing the first frost, while N21 and HOCP 96-540 lacked

similar capability.

The PIABS, which is a sensitive indicator of electron

transport efficiency (Kruger et al. 1997; Strasser et al. 2000;

Force et al. 2003; Strauss et al. 2007) were calculated from

the original O–J–I–P fluorescence transients. Compared to

the pre-frost state all varieties showed a decline in PIABS

following the first frost, however, N36 and LCP 85-384 then

maintained these values for at least a further 2 weeks longer

than N21 and HOCP 96-540 where further declines were

observed. Maintenance of electron transport efficiency for

at least 2 weeks longer indicates potential for cold accli-

mation in N36 and LCP 85-384 and corroborates the

observed recovery in O–J–I–P fluorescence rise kinetics.

The potential for cold acclimation in plants having the C4

photosynthetic pathway was recently demonstrated (Kakani

et al. 2008), and in sugarcane varieties the ability to cold

acclimate might be related to sensitivity of pyruvate,

orthophosphate dikinase (PPDK) and NADP-malate dehy-

drogenase (NADP-MDH) activity (Du et al. 1999). These

authors proposed that the inhibition of NADP-MDH and

PPDK activity limited CO2 assimilation capacity with

subsequent photoinhibition and upstream damage to the

electron transport systems. Sugarcane is a typical NADP-

ME subtype C4 plant and contains genes of both tropical

and sub-tropical species. It is therefore not surprising that

sugarcane is sensitive to low temperature, although differ-

ences in sensitivity between hybrid species have been

demonstrated (Du et al. 1999). These authors found large

differences in photosynthetic capacity between three hybrid

species during exposure to low temperature. In the hybrid

species that was least affected, almost all measured leaf

enzyme levels increased during cold exposure and photo-

synthetic rates also gradually increased by 20–30 % fol-

lowing the initial decline that occurred at the start of

exposure (Du et al. 1999). In another study sugarcane plants

grown at low temperature for extended periods of time also

showed some acclimation of the photosynthetic apparatus

(Ebrahim et al. 1998). These previous findings suggest that

varying degrees of cold acclimation potential might be

present in sugarcane. The results presented in the present

paper are to the best of our knowledge the first report of cold

acclimation ability under field conditions in sugarcane.

These previous findings, and the results presented here,

suggest that electron transport capacity is a main target

Variety

N36

LCP85-38

4

HOCP96-5

40 N21

ER

C c

on

ten

t(%

)

0

2

4

6

8

10Pre-frostHarvest

26% 20%

11%

8%

a

b

a

b

a

a

a a

Fig. 4 Stalk ERC content (%) in the four varieties pre-frost (black

bars) and at harvest (grey bars). The % values indicates how much

higher the ERC content was at harvest, relative to the pre-frost state,

and different lowercase letters indicate significant differences

(p \ 0.05) between the two sampling points for each variety. Error

bars indicate least significant difference at p \ 0.05 (LSD0.05)

Variety

N21 HOCP96-540 LCP85-384 N36

ER

C y

ield

(t/h

a)

0

1

2

3

4

5

6

a

bb

b

Fig. 5 ERC yields (t/ha) in the four varieties at harvest. The error

bar indicates the least significant difference (LSD) at p \ 0.05.

Different lowercase letters indicate significant differences (p \ 0.05)

between the varieties

186 Plant Growth Regul (2014) 72:181–187

123

during low temperature stress in sugarcane and that chlo-

rophyll a fluorescence is a sensitive tool for probing the

degree of damage and acclimation responses in sugarcane.

The observed cold acclimation in N36 and LCP 85-384

would have enabled maintenance of higher photosynthetic

efficiency (as indicated by the PIABS values), and thus also

ERC accumulation in the stalks for longer following the

first frost event. The results that showed that N36 and LCP

85-384 increased ERC content (%) by 26 and 20 %

between the pre-frost state and harvest, compared to N21

and HOCP 96-540 where the increase were only 8 and

11 %, supports this suggestion. This would have contrib-

uted towards N36 having a 30 % higher ERC yield (t/ha)

than N21 as well as the known yield advantage of HOCP

96-540 over LCP 85-384 (Dufrene and Tew 2004) being

reversed following frost.

In conclusion, the ability to maintain high ERC accu-

mulation capacity for longer following frost due to cold

acclimation could be an important factor determining

sugarcane yield performance in frost-prone areas. In

addition, O–J–I–P fluorescence rise kinetics show promise

as a rapid screening tool for assessment of cold acclimation

potential and frost tolerance in sugarcane.

Acknowledgments The author acknowledges Mr. Rudolph Koch

for provision of land and managing of the field trial on his farm Green

Hill (Kwazulu-Natal, South Africa).

References

Berry J, Bjorkman O (1980) Photosynthetic response and adaptation to

temperature in higher plants. Ann Rev Plant Physiol 31:491–543

Du YC, Nose A, Wasano K (1999) Effects of chilling temperature on

photosynthetic rates, photosynthetic enzyme activities and

metabolite levels in leaves of three sugarcane species. Plant,

Cell Environ 22:317–324

Dufrene EO, Tew TL (2004) HOCP 96-540, a chip off the old block?

Am Soc Sugar Cane Technol 24:82

Ebrahim MKH, Vogg G, Osman MNEH, Komor E (1998) Photo-

synthetic performance and adaptation of sugarcane at suboptimal

temperatures. J Plant Physiol 153:587–592

Edwards GE, Nakamoto H, Burnell JN, Hatch MD (1985) Pyruvate,

Pi dikinase and NADP-malate dehydrogenase in C4 photosyn-

thesis: properties and mechanism of light/dark regulation. Ann

Rev Plant Physiol 36:255–286

Eggleston G, Legendre B, Tew T (2004) Indicators of freeze-damaged

sugarcane varieties which can predict processing problems. Food

Chem 87:119–133

Force L, Critchley C, Van Rensen JJS (2003) New fluorescence parameters

for monitoring photosynthesis in plants. Photosynth Res 78:17–33

Haldimann P, Strasser RJ (1999) Effects of anaerobiosis as probed by

the polyphasic chlorophyll a fluorescence rise kinetic in pea

(Pisum sativum L.). Photosynth Res 62:67–83

Kakani VG, Boote KJ, Reddy KR, Lang DJ (2008) Response of

bahiagrass carbon assimilation and photosystem activity to

below optimum temperatures. Funct Plant Biol 35:1243–1254

Kautsky H, Hirsch A (1931) Neue versuche zur kohlensaure

assimilation. Naturwissenschaften 19:964

Kruger GHJ, Tsimilli-Michael M, Strasser RJ (1997) Light stress

provokes plastic and elastic modifications in structure and

function of photosystem II in camellia leaves. Physiol Plant

101:265–277

Lavorel J, Etienne AL (1977) In vivo chlorophyll fluorescence. Top

Photosynth 2:203–268

Lazar D, Ilık P (1997) High-temperature induced chlorophyll

fluorescence changes in barley leaves: comparison of the critical

temperatures determined from fluorescence induction and from

fluorescence temperature curve. Plant Sci 124:159–164

Osborne CP, Wythe EJ, Ibrahim DG, Gilbert ME, Ripley BS (2008)

Low temperature effects on leaf physiology and survivorship in

the C3 and C4 subspecies of Alloteropsis semialata. J Exp Bot

59:1743–1754

Papageorgio J (1975) Chlorophyll fluorescence: an intrinsic probe of

photosynthesis. In: Govindjee (ed) Bioenergetics of photosyn-

thesis. Academic Press, New York, pp 319–371

Rapacz M (2007) Chlorophyll a fluorescence transient during freezing

and recovery in winter wheat. Photosynthetica 45:409–418

Rapacz M, Sasal M, Gut M (2011) Chlorophyll fluorescence-based

studies of frost damage and the tolerance for cold-induced

photoinhibition in freezing tolerance analysis of Triticale

(9 Triticosecale Wittmack). J Agron Crop Sci 197:378–389

Rowley JA (1976) Development of freezing tolerance in leaves of C4

grasses. Aust J Plant Physiol 3:597–603

Rowley JA, Tunnicliffe CG, Taylor AO (1975) Freezing sensitivity of

leaf tissue of C4 grasses. Aust J Plant Physiol 2:447–451

Sage RF, Kubien DS (2007) The temperature response of C3 and C4

photosynthesis. Plant, Cell Environ 30:1086–1106

Snyman SJ, Meyer GM, Banasiak M, Nicholson TL, van Antwerpen

T, Naidoo P, Erasmus JD (2008) Micropropagation of sugarcane

via NovaCane�: preliminary steps in commercial application.

Proc S Afr Sugar Technol Assoc 81:513–516

Strasser RJ, Govindjee (1992) The F0 and the O–J–I–P fluorescence

rise in higher plants and algae. In: Argyroudi-Akoyunoglou JH

(ed) Regulation of chloroplast biogenesis. Plenum Press,

New York, USA, pp 423–426

Strasser BJ, Strasser RJ (1995) Measuring fast fluorescence transients

to address environmental questions: the JIP-test. In: Mathis P

(ed) Photosynthesis: from light to biosphere, vol V. Kluwer

Academic Publishers, The Netherlands, pp 977–980

Strasser RJ, Srivastava A, Govindjee (1995) Polyphasic chlorophyll

a fluorescence transient in plants and cyanobacteria. Photochem

Photobiol 61:32–42

Strasser RJ, Srivastava A, Tsimilli-Michael M (2000) The fluorescent

transient as a tool to characterise and screen photosynthesic

samples. In: Yunus M, Pathre U, Mohanty P (eds) Probing

photosynthesis: mechanisms, regulation and adaptation. Taylor

and Francis, London, pp 445–483

Strauss AJ, Kruger GHJ, Strasser RJ, Van Heerden PDR (2007) The

role of low soil temperature in the inhibition of growth and PSII

function during dark chilling in soybean genotypes of contrasting

tolerance. Physiol Plant 131:89–105

Trischuk RG, Schilling BS, Wisniewski M, Gusta LV (2006) Freezing

stress: systems biology to study cold tolerance. In: Madhava Rao

KV, Raghavendra AS, Janardhan Reddy K (eds) Physiology and

molecular biology of stress tolerance in plants. Springer,

Dordrecht, pp 131–155

Tsimilli-Michael M, Pecheux M, Strasser RJ (1999) Light and heat

stress adaptation of the symbionts of coral reef and temperate

foraminifers probed in hospite by the chlorophyll a fluorescence

kinetics O–J–I–P. Naturforsch 54C:671–680

Van Rensburg L, Kruger GHJ, Eggenberg P, Strasser RJ (1996) Can

screening criteria for drought resistance in Nicotiana tabacum L.

be derived from the polyphasic rise of the chlorophyll a fluores-

cence transient (OJIP)? S Afr J Bot 62:337–341

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